Extractability and chromatographic separation of proteins from potato (Solanum tuberosum L.) trimmings.

By-products from the potato processing industry, like potato trimmings, are sustainable sources of proteins. Here, a size-exclusion high performance liquid chromatography (SE-HPLC) method was applied to simultaneously determine the extractability and aggregation state of proteins from three batches of potato trimmings of different cultivars. Obtained SE-HPLC profiles allowed distinguishing between the patatin and protease inhibitor fractions of potato proteins. Moreover, only 75% of the crude proteins could be extracted in phosphate buffer containing sodium dodecyl sulfate and a reducing agent, indicating the presence of physical extraction barriers. Ball milling for 5 min significantly increased protein extractability, but prolonged treatment resulted in aggregation of native patatin and a reduced protein extractability. Microwave-dried trimmings had a lower protein extractability than freeze-dried trimmings. In future research, the SE-HPLC method can be used to examine changes in potato protein (fractions) as a result of processing.

Varying precipitation conditions allow directing the composition and physical properties of soy protein concentrates.

Soy protein concentrates (SPCs) are common food ingredients. They typically contain 65% (w/w) protein and ∼30% (w/w) carbohydrate. SPCs can be obtained with various protein precipitation conditions. A systematic study of the impact of these different protein precipitation protocols on the SPC protein composition and physical properties is still lacking. Here, SPCs were prepared via three different protocols, that is, isoelectric (pH 3.5-5.5), aqueous ethanol (50%-70% [v/v]), and Ca2+ ion (5-50 mM) based precipitations, and analyzed for (protein) composition, protein thermal properties, dispersibility, and water-holding capacity. SPCs precipitated at pH 5.5 or by adding 15 mM Ca2+ ions had a lower 7S/11S globulin ratio (∼0.40) than that (∼0.50) of all other SPC samples. Protein in SPCs obtained by isoelectric precipitation denatured at a significantly higher temperature than those in ethanol- or Ca2+ -precipitated SPCs. Precipitation with 50%-60% (v/v) ethanol resulted in pronounced denaturation of 2S albumin and 7S globulin fractions in SPCs. Additionally, increasing the precipitation pH from 3.5 to 5.5 and increasing the Ca2+ ion concentration from 15 to 50 mM caused a strong decrease of both the dispersibility of the protein in SPC and its water-holding capacity at pH 7.0. In conclusion, this study demonstrates that the SPC production process can be directed to obtain ingredients with versatile protein physicochemical properties toward potential food applications. PRACTICAL APPLICATION: This study demonstrates that applying different protein precipitation protocols allows obtaining SPCs that vary widely in (protein) composition and physical properties (such as protein dispersibility and water-holding capacity). These varying traits can greatly influence the suitability of SPCs as functional ingredients for specific applications, such as the production of food foams, emulsions, gels, and plant-based meat alternatives. The generated knowledge may allow targeted production of SPCs for specific applications.

Sucrose substitution in cake systems is not a piece of cake.

Successful sucrose replacement in cake systems requires thorough understanding of its functionality. Time-domain 1H NMR showed that water in the viscous aqueous phase isolated from cake batter by ultracentrifugation [i.e. the batter liquor (BL)] exhibits low mobility by its low T2 relaxation time (T2,D RT). This is due to its interactions with sucrose or sucrose replacers. The T2,D RT itself is positively related with the effective volumetric hydrogen bond density of sucrose or sucrose replacers. Sucrose additionally co-determines the quantity and viscosity of cake BL and thereby how much air the batter contains at the end of mixing. Like sucrose, maltitol and oligofructose provide adequate volumes of BL with low water mobility and thus sufficient air in the batter, while the rather insoluble mannitol and inulin do not. Differential scanning calorimetry and rapid viscosity analysis revealed, however, that, in contrast to sucrose and maltitol, oligofructose fails to provide appropriate timings of starch gelatinisation and protein denaturation, resulting in poor cake texture. The shortcomings of mannitol and oligofructose in terms of respectively ensuring appropriate gas content in batter and biopolymer transitions during baking can be overcome by using mixtures thereof. This work shows that successful sucrose substitutes or substitute mixtures must provide sufficient BL with low water mobility and ensure appropriate timings of starch and protein biopolymer transitions during baking.

Impact of process parameters on the specific volume of wholemeal wheat bread made using sourdough- and baker's yeast-based leavening strategies.

The final quality of wholemeal wheat bread is determined by the process parameter settings and leavening strategy. We hypothesise that the used leavening strategy may influence the optimal process parameter settings and, as such, the specific volume of the bread loaf. To analyse this interaction, bread was leavened with (i) a type 1 sourdough (SB), (ii) a type 1 sourdough combined with baker's yeast (YSB), or (iii) baker's yeast (YB). For each leavening strategy, the specific volume of bread, in response to variations in mixing time (4-10/4-14 min), water absorption (60-85 %), and proofing time (1-7/1-3 h), was analysed using an I-optimal response surface experimental design. Data modelling identified a substantially lower maximal specific volume of SB (2.13 mL/g), compared to YSB (3.30 mL/g) and YB (3.26 mL/g). The proofing time and water absorption mostly influenced the specific volume of the SB and YSB, respectively. However, the mixing and proofing times mainly affected the specific volume of YB. The type 1 sourdough reduced the mixing time and water absorption required for an optimal specific volume of bread compared to baker's yeast. These results challenge the idea of yielding higher volumes upon using sourdough compared to baker's yeast and highlight the importance of optimisation of bread dough formulations and breadmaking processes.

Study of the Fermentation Characteristics of Non-Conventional Yeast Strains in Sweet Dough.

Despite the diverse functions of yeast, only a relatively homogenous group of Saccharomyces cerevisiae yeasts is used in the baking industry. Much of the potential of the natural diversity of yeasts has not been explored, and the sensory complexity of fermented baked foods is limited. While research on non-conventional yeast strains in bread making is increasing, it is minimal for sweet fermented bakery products. In this study, the fermentation characteristics of 23 yeasts from the bakery, beer, wine, and spirits industries were investigated in sweet dough (14% added sucrose w/w dm flour). Significant differences in invertase activity, sugar consumption (0.78-5.25% w/w dm flour), and metabolite (0.33-3.01% CO2; 0.20-1.26% ethanol; 0.17-0.80% glycerol; 0.09-0.29% organic acids) and volatile compound production were observed. A strong positive correlation (R2 = 0.76, p < 0.001) between sugar consumption and metabolite production was measured. Several non-conventional yeast strains produced more positive aroma compounds and fewer off-flavors than the reference baker's yeast. This study shows the potential of non-conventional yeast strains in sweet dough.

The influence of varying levels of molecular oxygen on the functionality of azodicarbonamide and ascorbic acid during wheat bread making.

Air, and thus also molecular oxygen (O2), is incorporated in wheat flour dough during mixing. O2 participates in several (enzymatic) reactions, including those resulting in the oxidation of free sulfhydryl groups, thereby increasing dough strength and bread volume. We here incorporated different O2 levels in dough by mixing dough samples for a fixed time under different modified atmospheres which led to significant changes in dough free sulfhydryl contents and bread volumes. Although altering the mixing time not only impacted how much O2 was incorporated in dough but also the mechanical input, the changes in dough and bread properties when using different mixing times, largely depended on differences in O2 uptake. When used in bread recipes, redox agents such as azodicarbonamide (ADA) and ascorbic acid (AH2) impact the dough sulfhydryl contents and bread volumes. The effect of different levels of O2 incorporation on dough samples which contained ADA or AH2 was studied by altering the mixing time or the O2 content in the mixing atmosphere. Lower ADA levels were needed when dough was mixed under an atmosphere enriched in O2. As AH2 requires O2 to be converted to dehydroascorbic acid (DHA) to exert its improver effect, it came as a surprise that when it was included in a dough which was prepared under O2 enriched conditions, no additional impact was obtained and that, even under reduced O2 conditions, its use still resulted in an increased bread volume. These findings suggest that AH2 oxidase very effectively uses O2 to form DHA.

Sugar Levels Determine Fermentation Dynamics during Yeast Pastry Making and Its Impact on Dough and Product Characteristics.

Fermented pastry products are produced by fermenting and baking multi-layered dough. Increasing our knowledge of the impact of the fermentation process during pastry making could offer opportunities for improving the production process or end-product quality, whereas increasing our knowledge on the sugar release and consumption dynamics by yeast could help to design sugar reduction strategies. Therefore, this study investigates the impact of yeast fermentation and different sugar concentrations on pastry dough properties and product quality characteristics. First, yeasted pastry samples were made with 8% yeast and 14% sucrose on a wheat flour dry matter base and compared to non-yeasted samples. Analysis of saccharide concentrations revealed that sucrose was almost entirely degraded by invertase in yeasted samples after mixing. Fructans were also degraded extensively, but more slowly. At least 23.6 ± 2.6% of the released glucose was consumed during fermentation. CO2 production during fermentation contributed more to product height development than water and ethanol evaporation during baking. Yeast metabolites weakened the gluten network, causing a reduction in dough strength and extensibility. However, fermentation time had a more significant impact on dough rheology parameters than the presence of yeast. In balance, yeast fermentation did not significantly affect the calculated sweetness factor of the pastry product with 14% added sucrose. Increasing the sugar content (21%) led to higher osmotic stress, resulting in reduced sugar consumption, reduced CO2 and ethanol production and a lower product volume. A darker colour and a higher sweetness factor were obtained. Reducing the sugar content (7%) had the opposite effect. Eliminating sucrose from the recipe (0%) resulted in a shortened productive fermentation time due to sugar depletion. Dough rheology was affected to a limited extent by changes in sucrose addition, although no sucrose addition or a very high sucrose level (21%) reduced the maximum dough strength. Based on the insights obtained in this study, yeast-based strategies can be developed to improve the production and quality of fermented pastry.

Free wheat flour lipids decrease air-liquid interface stability in sponge cake batter.

The impact of free wheat flour lipids on air-liquid interface stability during sponge cake making was investigated. Therefore, the molecular population at the air-liquid interface in batters prepared with flour of which part of the lipids had been either relocated or removed prior to batter preparation was determined. Surface-active molecules were isolated from batter using a foam separation protocol. Diluted batter was whipped and the resulting foam was used as model system for the air-liquid interface in sponge cake batter. Relocating flour lipids prior to batter making enabled them to adsorb at the air-liquid interface in the foam. This limited the degree of protein adsorption at the air-liquid interface, but it did not impact the composition of the adsorbed protein population. Removing flour lipids prior to batter making resulted in foam containing relatively higher levels of lipids mainly originating from egg yolk. Prior removal of flour lipids impacted neither foam protein content nor foam protein composition. The resultant molecular population improved air-liquid interface stability in sponge cake batter. Thus, free wheat flour lipids and wheat flour lipids set free by solvent treatment decrease air-liquid interface stability in sponge cake batter mainly because they limit protein adsorption and, as such, interfere with the protein-dominated interface.

The Role of Intact and Disintegrated Egg Yolk Low-Density Lipoproteins during Sponge Cake Making and Their Impact on Starch and Protein Mediated Structure Setting.

The main sponge cake ingredients are flour, sucrose, eggs and leavening agents. Exogenous lipids (e.g., monoacylglycerols) are often used to increase air-liquid interface stability in the batter. There is a consumer trend to avoid foods containing such additives. We here reasoned that egg yolk may be an alternative source of surface-active lipids and set out to study the role of egg yolk lipids during sponge cake making. This was done by relocating or removing them prior to batter preparation using ethanol treatments and examining how this affects cake (batter) properties and structure setting during baking. Most egg yolk lipids occur within spherical low-density lipoproteins (LDLs) which were disintegrated by the ethanol treatments. Results showed that egg yolk lipids impact air-liquid interface stability and less so cake structure setting. To prepare high-quality sponge cakes by multistage mixing preferably intact LDLs or, alternatively, their components are needed to incorporate sufficient air during mixing and to stabilize it after mixing. It was also shown that the batter contains intact LDLs in the continuous phase and disintegrated LDLs at air-liquid interfaces. Sponge cake contains intact LDLs in the cake matrix, disintegrated LDLs at air-crumb interfaces and disintegrated LDLs incorporated into the protein network.

The role of exogenous lipids in starch and protein mediated sponge cake structure setting during baking.

While it is well established that using exogenous lipids (ELs) such as monoacylglycerols and polyglycerolesters of fatty acids improves gas cell incorporation and stability in sponge cake batter (SCB) and allows producing sponge cakes (SCs) with very high volume, fine grained crumb and soft texture, their impact on starch gelatinization and protein polymerization remained unknown. Here, differential scanning calorimetry and size-exclusion high performance liquid chromatography were performed on SC(B) samples prepared with or without ELs. Starch gelatinization and protein denaturation and polymerization started at temperatures exceeding 67 °C and mostly occurred up to a temperature of 96 °C. During further isothermal treatment at 96 °C the rigidity of the cake matrix (for which temperature-controlled time domain 1H NMR T2 relaxation times are a predictor) further increased mainly because of protein polymerization. While the temperature range of starch crystal melting was not affected by the use of ELs, protein polymerized more intensively in an 88 to 94 °C temperature range when SCB contained ELs. The more intense protein polymerization and the high water binding capacity of ELs presumably made the cake matrix more rigid at that point in time. The present results allow concluding that ELs not only impact air-liquid interface stability but also cake structure setting. Hence, both aspects most likely contribute to the superior quality of SCs containing ELs.

Wheat Gluten Amino Acid Analysis by High-Performance Anion-Exchange Chromatography with Integrated Pulsed Amperometric Detection.

The present chapter describes an accurate and user-friendly method for determining amino acid composition of wheat gluten proteins and their gliadin and glutenin fractions. The method consists of hydrolysis of the peptide bonds in 6.0 M hydrochloric acid (HCl) solution at 110 °C for 24 h, followed by evaporation of the acid and separation of the free amino acids by high-performance anion-exchange chromatography with integrated pulsed amperometric detection (HPAEC-IPAD). In contrast to conventional methods, the analysis requires neither pre- or post-column derivatization nor a time-consuming oxidation or derivatization step prior to hydrolysis. Correction factors account for incomplete release of Val and Ile even after hydrolysis for 24 h and for losses of Ser during evaporation. Gradient conditions including an extra eluent allow multiple sequential sample analyses without risk of Glu accumulation on the anion-exchange column which otherwise would result from high Gln levels in gluten proteins.

Impact of aqualysin 1 peptidase from Thermus aquaticus on molecular scale changes in the wheat gluten network during bread baking.

The impact of Aqualysin 1 (Aq1), the thermo-active peptidase of Thermus aquaticus, on wheat albumin, globulin, gliadin and glutenin proteins during heat treatment of wheat dough and bread baking was examined. The level of protein extractable in sodium dodecyl sulfate containing medium under non-reducing conditions (SDS-EP-NR) from wheat dough decreases upon heating to a lesser extent when Aq1 is used than in control experiments. The higher SDS-EP-NR level is caused by the release by Aq1 of peptides from the repetitive gluten protein domains during baking. These peptides are also extractable from bread crumb with salt solution. The resultant thermoset gluten network in bread crumb is mainly made up by protein from non-repetitive gluten domains.

Impact of chlorine treatment on properties of wheat flour and its components in the presence of sucrose.

Chlorine treatment increases the surface hydrophobicity of starch granules of soft wheat flour and reduces its gluten network forming capacity and apparent content of the flour lipids, the latter presumably by forming chlorinated derivatives. It also increases its solvent holding capacity as shown by proton nuclear magnetic resonance (1H NMR) experiments in presence of sucrose. The exchanging protons in the extra-granular space are present in an environment with restricted mobility, indicating strong interactions between chlorinated soft wheat flour (CSWF) components and solvent. Chlorination reduced the capability of free flour lipids to shield starch granules during rapid visco analysis (RVA). The higher viscosity reached in RVA with CSWF was attributed to not only its lipids having different properties but also its protein, resulting in improved starch swelling and amylose leaching. Finally, 1H NMR indicated that starch and protein networks in CSWF-sucrose gels were better structured than in gels prepared from non-chlorinated flour.

Wheat (Triticum aestivum L.) flour free lipid fractions negatively impact the quality of sponge cake.

The functionality of wheat flour lipids in sponge cakes prepared from flour, sugar, eggs and leavening agents only was investigated by altering their location or content in flour. Hexane (hex) or the more polar hexane:isopropanol (3:2 v/v) (hex:isoprop) were used to impact free flour lipid (FFL) or both FFL and bound flour lipid (BFL) fractions, respectively. Flour from which the FFLs were removed resulted in significantly improved cake volumes and crumb structures. Additional removal of part of the BFLs did not further impact cake quality. Prior contact of flour with hex:isoprop followed by gently removing the solvent broke native interactions between BFLs and starch or gluten and relocated more lipids than did hex. Cakes from flour with relocated lipids had coarse crumb structures. Our study demonstrates that FFLs and relocated flour lipids negatively impact sponge cake quality by disturbing air-liquid interface stabilization during mixing and the early phases of baking.

Ingredient Functionality During Foam-Type Cake Making: A Review.

Foam-type cakes are complex food systems. Their main ingredients are wheat flour, hen eggs, sugar, leavening agent, and, in some cases, oil and/or surfactants. In contrast to the vast amount of research outcomes on the contribution of ingredients to the quality of batter-type cake systems, information on the functionality and importance of the ingredients and their constituents in foam-type cake systems is lacking. This review defines foam-type cakes, describes how they are made, summarizes the current knowledge of factors determining their quality, and identifies the current knowledge gaps.

Conditions Governing Food Protein Amyloid Fibril Formation-Part I: Egg and Cereal Proteins.

Conditions including heating mode, time, temperature, pH, moisture and protein concentration, shear, and the presence of alcohols, chaotropic/reducing agents, enzymes, and/or salt influence amyloid fibril (AF) formation as they can affect the accessibility of amino acid sequences prone to aggregate. As some conditions applied on model protein resemble conditions in food processing unit operations, we here hypothesize that food processing can lead to formation of protein AFs with a compact cross ß-sheet structure. This paper reviews conditions and food constituents that affect amyloid fibrillation of egg and cereal proteins. While egg and cereal proteins often coexist in food products, their impact on each other's fibrillation remains unknown. Hen egg ovalbumin and lysozyme form AFs when subjected to moderate heating at acidic pH separately. AFs can also be formed at higher pH, especially in the presence of alcohols or chaotropic/reducing agents. Tryptic wheat gluten digests can form fibrillar structures at neutral pH and maize and rice proteins do so in aqueous ethanol or at acidic pH, respectively.

Conditions Governing Food Protein Amyloid Fibril Formation. Part II: Milk and Legume Proteins.

Both intrinsic and extrinsic factors impact amyloid formation of food proteins. We here review the impact of various conditions and food constituents on amyloid fibrillation of milk and legume proteins. Much less is known about casein and legume protein amyloid-like fibril formation than about that of whey proteins such as ß-lactoglobulin, α-lactalbumin, and bovine serum albumin. Proteins of both sources are often studied after heating under strong acidic (pH < 3) conditions. The latter induces changes in protein conformation and often peptide hydrolysis. At higher pH values, alcohols, chaotropic and/or reducing agents induce the conformational changes required to enhance fibrillation. Different types of food proteins can impact each other's fibrillation. Also, the presence of other food constituents can enhance or reduce it. No general conclusions on the mechanisms or impact of different food constituents on food proteins can be made. Optimal conditions for AF formation, that is, heating for several days at low pH, are rare in food processing. However, this does not exclude the possibility of AF formation in food products. For example, slow cooking of hydrolyzed proteins may enhance it. Future research should focus on the prevalence of AFs in complex food systems or model systems relevant for food processing.

Rational Design of Amyloid-Like Fibrillary Structures for Tailoring Food Protein Techno-Functionality and Their Potential Health Implications.

To control and enhance protein functionality is a major challenge for food scientists. In this context, research on food protein fibril formation, especially amyloid fibril formation, holds much promise. We here first provide a concise overview of conditions, which affect amyloid formation in food proteins. Particular attention is directed towards amyloid core regions because these sequences promote ordered aggregation. Better understanding of this process will be key to tailor the fibril formation process. Especially seeding, that is, adding preformed protein fibrils to protein solutions to accelerate fibril formation holds promise to tailor aggregation and fibril techno-functionality. Some studies have already indicated that food protein fibrillation indeed improves their techno-functionality. However, much more research is necessary to establish whether protein fibrils are useful in complex food systems and whether and to what extent they resist food processing unit operations. In this review the effect of amyloid formation on gelation, interfacial properties, foaming, and emulsification is discussed. Despite their prevalent role as functional structures, amyloids also receive a lot of attention due to their association with protein deposition diseases, prompting us to thoroughly investigate the potential health impact of amyloid-like aggregates in food. A literature review on the effect of the different stages of the human digestive process on amyloid toxicity leads us to conclude that food-derived amyloid fibrils (even those with potential pathogenic properties) very likely have minimal impact on human health. Nevertheless, prior to wide-spread application of the technology, it is highly advisable to further verify the lack of toxicity of food-derived amyloid fibrils.

Intact and Damaged Wheat Starch and Amylase Functionality During Multilayered Fermented Pastry Making.

The roles of native and damaged starch (DS) during fermented pastry making were examined by increasing the level of DS in wheat flour by ball-milling and/or by including amylase in the recipe. Increased DS levels increase laminated dough strength presumably by making less water available for the gluten. This effect was partly overcome by amylase use. During baking, a reduced resistance of the dough to gas cell expansion, as a result of enzymatic starch hydrolysis, seems responsible for increased pastry lift and improved crumb structure. Gelatinization of intact starch limits dough lift and expansion. Even at high amylase dosages structural collapse was limited, which suggests a significant role for gluten in pastry product structure formation. Differential scanning calorimetry and low-resolution 1 H nuclear magnetic resonance experiments indicated that increased levels of starch damage and amylase use impact the amylose network in the product and respectively increase and decrease the extent to which amylopectin retrogrades during storage. PRACTICAL APPLICATION: This research article evaluates the role of intact and damaged wheat starch during the production of fermented pastry products. An expanded knowledge on starch functionality during the different pastry production steps allows for a targeted selection of additives to improve product quality and production efficiency. The results obtained in this study can contribute to the realization of industrially feasible solutions for the production of quality pastry products.

Enzymatically Hydrolyzed Wheat Gluten as a Foaming Agent in Food: Incorporation in a Meringue Recipe as a Proof-of-Concept.

There is a growing interest in substituting animal proteins with plant protein sources in food systems. A notable example is the replacement of hen egg white (EW) protein, which is used in a wide range of food products because of its excellent foaming characteristics. Here, enzymatically hydrolyzed wheat gluten, which has greater solubility and better foaming properties than wheat gluten itself, was prepared and incorporated in a classical meringue recipe to investigate its potential as a foaming agent. Meringues based on gluten hydrolysates (GHs) had batters with lower density and greater apparent viscosity than those based solely on EW protein. Furthermore, after baking, these GH containing meringues had greater specific volume than those based on EW protein alone and no notable differences in color or texture between the different samples were noted. These outcomes were related to basic insights in the air-water interfacial behavior of GHs obtained in earlier studies. More specifically, the greater foaming capacity of GH than of EW protein solutions was related to their superior meringue batter (density and apparent viscosity) and product (specific volume) properties. While EW protein solutions had better foam stability than GH solutions (in the absence of sugar), this was apparently less relevant for meringue properties, probably due to the very high viscosity of the sugar rich batter, which could obscure differences in the intrinsic foam stabilizing ability of the samples. PRACTICAL APPLICATION: Replacing animal proteins with plant protein sources in the food industry is desirable from an economic and environmental perspective. Enzymatic hydrolysis serves as a tool to improve the foaming properties of water-insoluble wheat gluten proteins. We conclude that wheat gluten hydrolysates can be a valid functional alternative for egg white proteins in meringues, and possibly other food systems.